Rhodococcus ruber PTA-2, immobilization and applications thereof

A technology of PTA-2 and Rhodococcus rhodochrous, applied in the field of environmental microorganisms, can solve problems such as the impact on human health of the ecosystem, and achieve the effects of low production cost, good degradation effect and convenient use.

Active Publication Date: 2017-04-26
JIANGSU NANZI ENVIRONMENTAL PROTECTION SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Because of the strong hydrophilicity of the sulfonic acid group in its molecular structure, it is easily soluble in water, alcohol, and ether, but hardly soluble in benzene-based solvents such as benzene, toluene, and xylene; in additio...

Method used

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  • Rhodococcus ruber PTA-2, immobilization and applications thereof
  • Rhodococcus ruber PTA-2, immobilization and applications thereof
  • Rhodococcus ruber PTA-2, immobilization and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Isolation and Identification of Example 1 Rhodococcus Erythrococcus PTA-2

[0024] Rhodococcus erythrococcus PTA-2 was isolated and screened from the activated sludge pool of the sewage treatment station of the pesticide chemical plant.

[0025] The specific process of screening is:

[0026] 1) Take 5ml of activated sludge from the sewage treatment station of the pesticide chemical factory and inoculate it into the inorganic salt medium containing 200mg / L p-toluenesulfonic acid. The formula is: 200mg / L p-toluenesulfonic acid, 1.5g / L K 2 HPO 4 , 0.5g / L KH 2 PO 4 , 0.2g / L MgSO 4 ·7H 2 O, 1.0g / L NaCl, 1.0g / L (NH 4 ) 2 SO 4 After culturing in a shaker incubator at 30°C-37°C for 7 days, remove 5m of the mixed solution and inoculate it into an inorganic salt medium containing 400mg / L p-toluenesulfonic acid. The formula is: 400mg / L p-toluenesulfonic acid , 1.5g / L K 2 HPO 4 , 0.5g / L KH 2 PO 4 , 0.2g / L MgSO 4 ·7H 2 O, 1.0g / L NaCl, 1.0g / L (NH 4 ) 2 SO 4 , afte...

Embodiment 2

[0029] Embodiment 2 Rhodococcus erythrococcus PTA-2 takes p-toluenesulfonic acid as the culture status of sole carbon source

[0030] Pick a single colony from the plate where Rhodococcus rubrum PTA-2 grows, inoculate it in LB liquid medium, culture it in 30°C, 160rpm shaker for 12h (OD600≈1.0), centrifuge at 8000-12000rpm, discard the upper The supernatant was washed three times with sterile water, and resuspended in an equal volume of sterile water as the seed solution.

[0031] Fill a 250mL Erlenmeyer flask with an inorganic salt medium containing 200mg / L p-toluenesulfonic acid, the formulation of which is: 200mg / L p-toluenesulfonic acid, 1.5g / L K 2 HPO 4 , 0.5g / L KH 2 PO 4 , 0.2g / L MgSO 4 ·7H 2 O, 1.0g / L NaCl, 1.0g / L (NH 4 ) 2 SO 4 , insert the PTA-2 seed solution according to the inoculum size of 1% (volume fraction), cultivate in 30 ℃, 160rpm shaker, take out 4mL culture solution every 2h, discard the supernatant after the centrifuge centrifugation of sample throug...

Embodiment 3

[0032] Embodiment 3 Rhodococcus rubella PTA-2 is prepared into the method for immobilized pellet

[0033] 1) Use an inoculation loop to pick a small amount of bacteria on the slant medium and inoculate it in LB liquid medium, and culture it to the logarithmic phase at 30°C and 160r / min with shaking;

[0034] 2) Centrifuge the above cultured bacterial solution at 10000r / min for 2min, discard the supernatant, add the same volume of sterile water, shake well and centrifuge at 10000r / min for 2min, wash twice in this way, and use the same volume Suspended in sterile water to make a bacterial fluid.

[0035] 3) Weigh 1.2g of sodium alginate and dissolve it in 20ml of distilled water, add 500mg / L activated carbon powder, and disperse by ultrasonic.

[0036] 4) Take 1ml of the bacterial liquid and 20ml of the step 3) solution at room temperature and mix well, then use a syringe to drop the mixed solution into 4% CaCl at room temperature 2 The solution was cross-linked and calcified ...

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Abstract

The invention discloses Rhodococcus ruber PTA-2, which is classified and named Rhodococcus ruber PTA-2, is preserved in the China General Microbiological Culture Collection Center on February 9, 2015, and has the preservation number of CGMCC NO.10538. According to the present invention, the Rhodococcus ruber PTA-2 can grow by adopting p-toluenesulfonic acid as the sole carbon source, wherein the degradation rate on the 500 mg/L p-toluenesulfonic acid is 96%; the immobilized pellets prepared from the Rhodococcus ruber PTA-2 provides the good p-toluenesulfonic acid degradation effect, wherein the degradation rate on the 500 mg/L p-toluenesulfonic acid is more than 90%; and the production cost is low, the use is convenient, and the Rhodococcus ruber PTA-2 is suitable for promotion and application in the p-toluenesulfonic acid-containing wastewater treatment.

Description

technical field [0001] The invention relates to the technical field of environmental microorganisms, in particular to a rhodococcus rubella PTA-2 and its immobilization and application. Background technique [0002] p-toluenesulfonic acid is an important chemical raw material, widely used as a synthetic medicine (such as naproxen, doxycycline, etc.), pesticides (such as quizalofop-ethyl, dicofol, etc.) and dyes (acid orange 7, etc.) Intermediate; also used as a catalyst for the synthesis of esters (such as benzyl propionate, benzyl butyrate, etc.), a stabilizer for polymerization reactions, and a curing agent for resins, paints, coatings, casting, wood-based panels and other industries; at the same time in It is also widely used in petrochemical, medical, plastic and other industries. [0003] P-toluenesulfonic acid is moderately toxic and irritating to eyes, skin, and mucous membranes. Because of the strong hydrophilicity of the sulfonic acid group in its molecular struct...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N11/14C12N11/10C02F3/34C12R1/01C02F101/34
CPCC02F3/34C02F2101/40C12N11/10C12N11/14C12N1/205C12R2001/01
Inventor 李珊珊姜自茹林举媚张永乐
Owner JIANGSU NANZI ENVIRONMENTAL PROTECTION SCI & TECH
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